Use of tight junction antagonists in the treatment of acute lung injury and acute respiratory distress syndrome

ABSTRACT

The present application provides compositions and methods for treating acute lung injury and acute respiratory distress syndrome. The methods include administering one or more tight junction antagonists to the lung of a subject in need thereof.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.14/971,675, filed Dec. 16, 2015, which is a continuation of U.S. patentapplication Ser. No. 13/852,569, filed Mar. 28, 2013, which is acontinuation of U.S. patent application Ser. No. 12/667,192, which isthe U.S. national stage of PCT/US2008/068465, filed Jun. 27, 2008, whichclaims priority to and the benefit of U.S. Provisional PatentApplication No. 60/947,228, filed Jun. 29, 2007, the entire contents ofall of which are hereby incorporated by reference in their entireties.

BACKGROUND

Acute Respiratory Distress Syndrome (ARDS) presents in about 150,000individuals in the US annually, with a mortality rate of 30-50%. ARDSoccurs in response to diverse forms of severe injury, in which lungedema results in respiratory failure. The current standard of care forARDS is limited to the management of the disease through supportivemechanical ventilation. The loss of endothelial barrier integrity iscentral to the pulmonary edema that occurs in ARDS.

Triggering causes for Acute Lung Injury (ALI) including ARDS can, forexample, be diffuse pulmonary infections (e.g. due to viruses, bacteria,fungi), aspiration of liquids (e.g. gastric juice or water), inhalationof toxins or irritants (e.g. chlorine gas, nitrogen oxides, smoke),direct or indirect trauma (e.g. multiple fractures or pulmonarycontusion), systemic reactions to inflammations outside the lung (e.g.hemorrhagic pancreatitis, gram-negative septicemia), transfusions ofhigh blood volumes or alternatively after cardiopulmonary bypass.

The pulmonary vascular endothelium lines the intravascular space andpresents a selective barrier that actively regulates paracellularmovement of circulating fluid, macromolecules, and cells, intoextravascular tissues and compartments. Loss of this endothelial barrierintegrity is the central defect found in ALI and ARDS. The host responseto a wide range of injurious stimuli includes the biosynthesis andrelease of endogenous mediators, some of which can open the paracellularpathway in lung microvascular endothelia. Several of these mediatorshave been identified, including tumor necrosis factor α, interleukin-1,thrombospondin-1, and SPARC/osteonectin, and these mediators have beenestablished as factors that disrupt endothelial barrier integrity. Inrecent preliminary studies, we found that ΔG, the active domain ofzonula occludens toxin (ZOT) of Vibrio cholerae, increases paracellularpermeability across human lung microvascular endothelial cells(HMVEC-Ls).

ZOT and ΔG have been previously identified as tight junction agonists,i.e., compounds that mediate or facilitate or augment the physiological,transient opening of tight junctions that form a barrier betweenadjacent epithelial cells. The ability of ZOT and ΔG to open tightjunctions has been used to facilitate the transfer of macromoleculeacross epithelial barriers (see U.S. Pat. No. 5,665,389 and Salama etal. J. Pharmacology and Experimental Therapeutics 312(1):199-205, 2005).ZOT has been shown to act as a tight junction agonist that allowsopening of tight junctions between adjacent mucosal epithelial cells.Compounds that antagonize the opening of tight junctions have beenidentified (see U.S. Pat. Nos. 6,458,925, 6,670,448, 6,936,689 and7,189,696). One such antagonist is currently in Phase II clinical trialsfor the treatment of celiac disease where it protects against loss ofgut mucosal barrier function.

There remains a need in the art for compositions and methods for thetreatment of ALI and ARDS. These and other needs are met by the presentinvention.

SUMMARY OF THE INVENTION

The present invention provides compositions and methods for thetreatment of an excessive or undesirable permeability of lung tissuecontaining tight junctions. Methods of the invention may includeadministering to a subject in need thereof a composition comprising atight junction antagonist. In some embodiments, a subject is any mammal,for example, a human. In some embodiments, a tight junction antagonistmay be a peptide, for example, a peptide comprising a sequence selectedfrom the group consisting of SEQ ID NOs:1-24. In some embodiments, atight junction antagonist may be a peptide, for example, a peptidecomprising the sequence G-G-V-L-V-Q-P-G (SEQ ID NO:15). In someembodiments, a tight junction antagonist may consist of the sequenceG-G-V-L-V-Q-P-G (SEQ ID NO:15).

In some embodiments, the present invention provides compositions andmethods for the treatment of acute lung injury. Such methods maycomprise administering to a subject in need thereof a compositioncomprising a tight junction antagonist. In some embodiments, a subjectis any mammal, for example, a human. In some embodiments, a tightjunction antagonist may be a peptide, for example, a peptide comprisinga sequence selected from the group consisting of SEQ ID NOs:1-24. Insome embodiments, a tight junction antagonist may be a peptide, forexample, a peptide comprising the sequence G-G-V-L-V-Q-P-G (SEQ IDNO:15). In some embodiments, a tight junction antagonist may consist ofthe sequence G-G-V-L-V-Q-P-G (SEQ ID NO:15).

In some embodiments, the present invention provides materials and methodfor the treatment of acute respiratory distress syndrome. Such methodsmay include administering to a subject in need thereof a compositioncomprising a tight junction antagonist. In some embodiments, a subjectis any mammal, for example, a human. In some embodiments, a tightjunction antagonist may be a peptide, for example, a peptide comprisinga sequence selected from the group consisting of SEQ ID NOs:1-24. Insome embodiments, a tight junction antagonist may be a peptide, forexample, a peptide comprising the sequence G-G-V-L-V-Q-P-G (SEQ IDNO:15). In some embodiments, a tight junction antagonist may consist ofthe sequence G-G-V-L-V-Q-P-G (SEQ ID NO:15). Compositions for use in themethods of the invention may also comprise one or more additionalcomponents. Examples of additional components include, but are notlimited to, aminosalicylates, corticosteroids, immunomodulators,antibiotics, cytokines, chemokines and biologic therapeutics.Compositions for use in the methods of the invention may be formulatedin any manner known to those skilled in the art, for example, thecompositions may be formulated for pulmonary delivery.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic representation of the experimental protocol usedin the examples.

FIG. 2 is a bar graph showing lung permeability as a function of thetreatment regimen comparing IV versus IT administration of tightjunction antagonist.

FIG. 3 is a bar graph showing lung permeability as a function of theamount of tight junction antagonist applied with anti-BSA antibodyintratracheally.

DETAILED DESCRIPTION OF THE INVENTION

Antagonists of Tight Junction Opening

As used herein, tight junction antagonists prevent, inhibit or reducethe opening of tight junctions, for example, the opening of tightjunctions induced by a tight junction agonist. A tight junctionantagonist may bind to the receptor that mediates tight junction agonistinduced opening of tight junctions. For example, a tight junctionantagonist may bind to the ZOT receptor and prevent, inhibit, reduce orreverse the tight junction opening triggered by the tight junctionagonist ZOT.

As used herein a subject is any animal, e.g., mammal, upon which methodsof the invention may be practiced and/or to which materials of thepresent invention may be administered. Subjects include, but are notlimited to, humans.

Any antagonist of tight junction opening may be used in the practice ofthe present invention. For example, antagonists of the invention maycomprise peptide antagonists. Examples of peptide antagonists include,but are not limited to, peptides that comprise an amino acid sequenceselected from the group consisting of

(SEQ ID NO: 1) Gly Arg Val Cys Val Gln Pro Gly, (SEQ ID NO: 2)Gly Arg Val Cys Val Gln Asp Gly, (SEQ ID NO: 3)Gly Arg Val Leu Val Gln Pro Gly, (SEQ ID NO: 4)Gly Arg Val Leu Val Gln Asp Gly, (SEQ ID NO: 5)Gly Arg Leu Cys Val Gln Pro Gly, (SEQ ID NO: 6)Gly Arg Leu Cys Val Gln Asp Gly, (SEQ ID NO: 7)Gly Arg Leu Leu Val Gln Pro Gly, (SEQ ID NO: 8)Gly Arg Leu Leu Val Gln Asp Gly, (SEQ ID NO: 9)Gly Arg Gly Cys Val Gln Pro Gly, (SEQ ID NO: 10)Gly Arg Gly Cys Val Gln Asp Gly, (SEQ ID NO: 11)Gly Arg Gly Leu Val Gln Pro Gly, (SEQ ID NO: 12)Gly Arg Gly Leu Val Gln Asp Gly, (SEQ ID NO: 13)Gly Gly Val Cys Val Gln Pro Gly, (SEQ ID NO: 14)Gly Gly Val Cys Val Gln Asp Gly, (SEQ ID NO: 15)Gly Gly Val Leu Val Gln Pro Gly, (SEQ ID NO: 16)Gly Gly Val Leu Val Gln Asp Gly, (SEQ ID NO: 17)Gly Gly Leu Cys Val Gln Pro Gly, (SEQ ID NO: 18)Gly Gly Leu Cys Val Gln Asp Gly, (SEQ ID NO: 19)Gly Gly Leu Leu Val Gln Pro Gly, (SEQ ID NO: 20)Gly Gly Leu Leu Val Gln Asp Gly, (SEQ ID NO: 21)Gly Gly Gly Cys Val Gln Pro Gly, (SEQ ID NO: 22)Gly Gly Gly Cys Val Gln Asp Gly, (SEQ ID NO: 23)Gly Gly Gly Leu Val Gln Pro Gly,  and (SEQ ID NO: 24)Gly Gly Gly Leu Val Gln Asp Gly 

Examples of peptide antagonists include, but are not limited to,peptides that consist of an amino acid sequence selected from the groupconsisting of SEQ ID NOs:1-24. Examples of peptide antagonists of tightjunctions can be found in U.S. Pat. Nos. 6,458,925, 6,670,448,6,936,689, and 7,189,696 the entire contents of which are specificallyincorporate herein by reference.

When the antagonist is a peptide, any length of peptide may be used.Generally, the size of the peptide antagonist will range from about 6 toabout 100, from about 6 to about 90, from about 6 to about 80, fromabout 6 to about 70, from about 6 to about 60, from about 6 to about 50,from about 6 to about 40, from about 6 to about 30, from about 6 toabout 25, from about 6 to about 20, from about 6 to about 15, from about6 to about 14, from about 6 to about 13, from about 6 to about 12, fromabout 6 to about 11, from about 6 to about 10, from about 6 to about 9,or from about 6 to about 8 amino acids in length. Peptide antagonists ofthe invention may be from about 8 to about 100, from about 8 to about90, from about 8 to about 80, from about 8 to about 70, from about 8 toabout 60, from about 8 to about 50, from about 8 to about 40, from about8 to about 30, from about 8 to about 25, from about 8 to about 20, fromabout 8 to about 15, from about 8 to about 14, from about 8 to about 13,from about 8 to about 12, from about 8 to about 11, or from about 8 toabout 10 amino acids in length. Peptide antagonists of the invention maybe from about 10 to about 100, from about 10 to about 90, from about 10to about 80, from about 10 to about 70, from about 10 to about 60, fromabout 10 to about 50, from about 10 to about 40, from about 10 to about30, from about 10 to about 25, from about 10 to about 20, from about 10to about 15, from about 10 to about 14, from about 10 to about 13, orfrom about 10 to about 12 amino acids in length. Peptide antagonists ofthe invention may be from about 12 to about 100, from about 12 to about90, from about 12 to about 80, from about 12 to about 70, from about 12to about 60, from about 12 to about 50, from about 12 to about 40, fromabout 12 to about 30, from about 12 to about 25, from about 12 to about20, from about 12 to about 15, or from about 12 to about 14 amino acidsin length. Peptide antagonists of the invention may be from about 15 toabout 100, from about 15 to about 90, from about 15 to about 80, fromabout 15 to about 70, from about 15 to about 60, from about 15 to about50, from about 15 to about 40, from about 15 to about 30, from about 15to about 25, from about 15 to about 20, from about 19 to about 15, fromabout 15 to about 18, or from about 17 to about 15 amino acids inlength.

The peptide antagonists can be chemically synthesized and purified usingwell-known techniques, such as described in High Performance LiquidChromatography of Peptides and Proteins: Separation Analysis andConformation, Eds. Mant et al., C.R.C. Press (1991), and a peptidesynthesizer, such as Symphony (Protein Technologies, Inc); or by usingrecombinant DNA techniques, i.e., where the nucleotide sequence encodingthe peptide is inserted in an appropriate expression vector, e.g., an E.coli or yeast expression vector, expressed in the respective host cell,and purified therefrom using well-known techniques.

Compositions

Typically, compositions, such as pharmaceutical compositions, comprisinga tight junction antagonist (e.g., peptide tight junction antagonist)comprise a pharmaceutically effective amount of the antagonist. Thepharmaceutically effective amount of antagonist (e.g., peptide tightjunction antagonist) employed in any given composition may varyaccording to factors such as the disease state, age, sex, and weight ofthe individual. Dosage regimens may be adjusted to provide the optimumtherapeutic response. For example, a single bolus may be administered,several divided doses may be administered over time or the dose may beproportionally reduced or increased as indicated by the exigencies ofthe therapeutic situation.

Generally, the amount of antagonist used for preventing, amelioratingand/or treating a disease in a subject will be in the range of about 1.0μg to 1 g, preferably about 1 mg to about 1000 mg, or from about 10 mgto about 100 mg, or from about 10 mg to about 50 mg, or from about 10 mgto about 25 mg of antagonist per dose.

Compositions of the invention may comprise one or more tight junctionantagonists at a level of from about 0.1 wt % to about 20 wt %, fromabout 0.1 wt % to about 18 wt %, from about 0.1 wt % to about 16 wt %,from about 0.1 wt % to about 14 wt %, from about 0.1 wt % to about 12 wt%, from about 0.1 wt % to about 10 wt %, from about 0.1 wt % to about 8wt %, from about 0.1 wt % to about 6 wt %, from about 0.1 wt % to about4 wt %, from about 0.1 wt % to about 2 wt %, from about 0.1 wt % toabout 1 wt %, from about 0.1 wt % to about 0.9 wt %, from about 0.1 wt %to about 0.8 wt %, from about 0.1 wt % to about 0.7 wt %, from about 0.1wt % to about 0.6 wt %, from about 0.1 wt % to about 0.5 wt %, fromabout 0.1 wt % to about 0.4 wt %, from about 0.1 wt % to about 0.3 wt %,or from about 0.1 wt % to about 0.2 wt % of the total weight of thecomposition. Compositions of the invention may comprise one or moretight junction antagonists at a level of about 0.1 wt %, about 0.2 wt %,about 0.3 wt %, about 0.4 wt %, about 0.5 wt %, about 0.6 wt %, about0.7 wt %, about 0.8 wt %, or about 0.9 wt % based on the total weight ofthe composition.

Compositions of the invention may comprise one or more tight junctionantagonists at a level of from about 1 wt % to about 20 wt %, from about1 wt % to about 18 wt %, from about 1 wt % to about 16 wt %, from about1 wt % to about 14 wt %, from about 1 wt % to about 12 wt %, from about1 wt % to about 10 wt %, from about 1 wt % to about 9 wt %, from about 1wt % to about 8 wt %, from about 1 wt % to about 7 wt %, from about 1 wt% to about 6 wt %, from about 1 wt % to about 5 wt %, from about 1 wt %to about 4 wt %, from about 1 wt % to about 3 wt %, or from about 1 wt %to about 2 wt % of the total weight of the composition. Compositions ofthe invention may comprise one or more tight junction effectors at alevel of about 1 wt %, about 2 wt %, about 3 wt %, about 4 wt %, about 5wt %, about 6 wt %, about 7 wt %, about 8 wt %, or about 9 wt % based onthe total weight of the composition.

Compositions of the invention may be formulated for pulmonary delivery(e.g., may be pulmonary dosage forms). Typically such compositions maybe provided as pharmaceutical aerosols, e.g., solution aerosols orpowder aerosols. Those of skill in the art are aware of many differentmethods and devices for the formation of pharmaceutical aerosols, forexample, those disclosed by Sciarra and Sciarra, Aerosols, in Remington:The Science and Practice of Pharmacy, 20th Ed., Chapter 50, Gennaro etal. Eds., Lippincott, Williams and Wilkins Publishing Co., (2000).

In one embodiment, the dosage forms are in the form of a powder aerosol(i.e, comprise particles). These are particularly suitable for use ininhalation delivery systems. Powders may comprise particles of any sizesuitable for administration to the lung.

Powder formulations may optionally contain at least one particulatepharmaceutically acceptable carrier known to those of skill in the art.Examples of suitable pharmaceutical carriers include, but are notlimited to, saccharides, including monosaccharides, disaccharides,polysaccharides and sugar alcohols such as arabinose, glucose, fructose,ribose, mannose, sucrose, trehalose, lactose, maltose, starches,dextran, mannitol or sorbitol. In one embodiment, a powder formulationmay comprise lactose as a carrier.

Powder formulations may be contained in any container known to those inthe art. Containers may be capsules of, for example, gelatin or plastic,or in blisters (e.g. of aluminum or plastic), for use in a dry powderinhalation device. In some embodiments, the total weight of theformulation in the container may be from about 5 mg to about 50 mg. Inother embodiments, powder formulations may be contained in a reservoirin a multi-dose dry powder inhalation device adapted to deliver asuitable amount per actuation.

Powder formulations typically comprise small particles. Suitableparticles can be prepared using any means known in the art, for example,by grinding in an airjet mill, ball mill or vibrator mill, sieving,microprecipitation, spray-drying, lyophilisation or controlledcrystallisation. Typically, particles will be about 10 microns or lessin diameter. Particles for use in the compositions of the invention mayhave a diameter of from about 0.1 microns to about 10 microns, fromabout 0.1 microns to about 9 microns, from about 0.1 microns to about 8microns, from about 0.1 microns to about 7 microns, from about 0.1microns to about 6 microns, from about 0.1 microns to about 5 microns,from about 0.1 microns to about 4 microns, from about 0.1 microns toabout 3 microns, from about 0.1 microns to about 2 microns, from about0.1 microns to about 1 micron, from about 0.1 microns to about 0.5microns, from about 1 micron to about 10 microns, from about 1 micron toabout 9 microns, from about 1 micron to about 8 microns, from about 1micron to about 7 microns, from about 0.1 micron to about 6 microns,from about 1 micron to about 5 microns, from about 1 micron to about 4microns, from about 1 micron to about 3 microns, from about 1 micron toabout 2 microns, from about 2 microns to about 10 microns, from about 2microns to about 9 microns, from about 2 microns to about 8 microns,from about 2 microns to about 7 microns, from about 2 microns to about 6microns, from about 2 microns to about 5 microns, from about 2 micronsto about 4 microns, or from about 2 microns to about 3 microns. In someembodiments, particles for use in the invention may be about 1 micron,about 2 microns, about 3 microns, about 4 microns, about 5 microns,about 6 microns, about 7 microns, about 8 microns, about 9 microns, orabout 10 microns in diameter.

In one embodiment, the dosage forms are in the form of a solutionaerosol (i.e., comprise droplets). Typically, droplets will be about 10microns or less in diameter. Droplets for use in the compositions of theinvention may have a diameter of from about 0.1 microns to about 10microns, from about 0.1 microns to about 9 microns, from about 0.1microns to about 8 microns, from about 0.1 microns to about 7 microns,from about 0.1 microns to about 6 microns, from about 0.1 microns toabout 5 microns, from about 0.1 microns to about 4 microns, from about0.1 microns to about 3 microns, from about 0.1 microns to about 2microns, from about 0.1 microns to about 1 micron, from about 0.1microns to about 0.5 microns, from about 1 micron to about 10 microns,from about 1 micron to about 9 microns, from about 1 micron to about 8microns, from about 1 micron to about 7 microns, from about 1 micron toabout 6 microns, from about 1 micron to about 5 microns, from about 1micron to about 4 microns, from about 1 micron to about 3 microns, fromabout 1 micron to about 2 microns, from about 2 microns to about 10microns, from about 2 microns to about 9 microns, from about 2 micronsto about 8 microns, from about 2 microns to about 7 microns, from about2 microns to about 6 microns, from about 2 microns to about 5 microns,from about 2 microns to about 4 microns, or from about 2 microns toabout 3 microns. In some embodiments, particles and/or droplets for usein the invention may be about 1 micron, about 2 microns, about 3microns, about 4 microns, about 5 microns, about 6 microns, about 7microns, about 8 microns, about 9 microns, or about 10 microns indiameter.

In addition to a tight junction antagonist, compositions of theinvention may further comprise one or more additional therapeuticagents, particularly therapeutic agents conventionally used for treatinglung conditions such as ALI and ARDS. Such additional therapeutic agentsinclude, but are not limited to, steroids and other anti-inflammatorycompounds. Suitable therapeutic agents may include one or more ofaminosalicylates, corticosteroids, immunomodulators, antibiotics,cytokines, chemokines and biologic therapies.

Compositions of the invention may also comprise one or morepharmaceutically acceptable excipients. Suitable excipients include, butare not limited to, buffers, buffer salts, bulking agents, salts,surface active agents, acids, bases, sugars, and binders.

Methods of Use

The compositions of the invention can be used for preventing, slowingthe onset of, ameliorating and/or treating ALI and ARDS. Typically,compositions may be administered one or more times each day in an amountsuitable to prevent, reduce the likelihood of an attack of, or reducethe severity of an attack of the underlying disease condition.

In some embodiments, compositions of the invention may be givenrepeatedly over a protracted period, i.e., may be chronicallyadministered. Typically, compositions may be administered one or moretimes each day in an amount suitable to treat ALI and ARDS.

In some embodiments, compositions of the invention may be used to treatacute symptoms of ALI and ARDS. Typically, embodiments of this type willrequire administration of the compositions of the invention to a subjectundergoing an acute episode in an amount suitable to reduce the severityof the symptoms. One or more administration may be used.

A composition according to the present invention may be pre-mixed priorto administration, or can be formed in vivo when two or more components(e.g., a tight junction antagonist and an additional therapeutic agent)are administered within 24 hours of each other. When administeredseparately, the components may be administered in either order (e.g.tight junction antagonist first followed by additional therapeutic agentor additional therapeutic agent first followed by tight junctionantagonist). The components can be administered within a time span ofabout 12 hours, about 8 hours, about 4 hours, about 2 hours, about 1hour, about 0.5 hour, about 0.25 hour, about 0.1 hour, about 1 minute,about 0.5 minute, or about 0.1 minute.

Administration of the compositions described above, e.g., compositionscomprising one or more tight junction antagonists and optionallycomprising one or more additional therapeutic agents, may be byinhalation. For example, one or more tight junction antagonists and oneor more additional therapeutic agents or a mixture thereof, may be ininhalable form. An example of an inhalable form is an atomizablecomposition such as an aerosol comprising the tight junction antagonist,either alone or in combination with one or more additional therapeuticagents, in solution or dispersion in a propellant, or a nebulizablecomposition comprising a solution or dispersion of the active ingredientin an aqueous, organic or aqueous/organic medium. For example, theinhalable form of the compositions of the invention may be an aerosolcomprising a mixture of one or more tight junction antagonists and oneor more additional therapeutic agents in solution or dispersion in apropellant, or a combination of an aerosol containing one or more tightjunction antagonists in solution or dispersion in a propellant with anaerosol containing one or more therapeutic agents in solution ordispersion in a propellant. In another example, the inhalable form ofthe compositions of the invention my be a nebulizable compositioncomprising a dispersion of one or more tight junction antagonists andone or more additional therapeutic agents in an aqueous, organic oraqueous/organic medium, or a combination of a dispersion of one or moretight junction antagonists with a dispersion of one or more additionaltherapeutic agents in such a medium.

Compositions of the invention may be administered in conjunction withone or more additional therapeutic agents, particularly therapeuticagents used conventionally for treating lung conditions such as ALI andARDS. Such additional therapeutic agents include, but are not limitedto, steroids and other anti-inflammatory compounds. Suitable therapeuticagents may include one or more of aminosalicylates, corticosteroids,immunomodulators, antibiotics, cytokines, chemokines and biologictherapies.

Compositions of the invention and one or more additional therapeuticagents may be administered simultaneously, or alternatively compositionsof the invention and one or more additional therapeutic agents may notbe administered simultaneously. Furthermore, compositions of theinvention may be administered prior to administration of one or moreadditional therapeutic agents, or alternatively compositions of theinvention may be administered subsequent to administration of one ormore additional therapeutic agents.

The following examples are provided for illustrative purposes only, andare in no way intended to limit the scope of the present invention.

EXAMPLES

The IgG immune complex model is a well established model of lung injuryand is shown schematically in FIG. 1. Briefly, a heterologous antibodymix containing antibodies to a known antigen are injected into an animalintravenously (IV) or intratracheally (IT). The known antigen and asmall amount of radiolabelled known antigen are injected into the animalIV. This results in immune complex (IC) formation between the antigenand the cognate antibodies in the heterologous antibody mix. The immunecomplex binds to binds to the Fe gamma receptor (FcγR) and thisinitiates an inflammatory cascade and leads to injury. One of theresults of the inflammatory cascade is an increase in lung permeabilitythat increases with extent of injury. The increase in lung permeabilityis quantified by measuring the radiolabelled antigen present in lungversus blood where radiolabel in the lung versus blood increases withpermeability. (See Johnson and Ward, J. Clin. Investigation 54:349-357,1974).

FIG. 2 shows the results of an experiment where 4-5 mice per arm weretreated as described above where the known antigen was bovine serumalbumin (BSA) the heterologous antibody mix included antibody to bovineserum albumin (anti-BSA) and the treatment was carried out in thepresence and absence of tight junction antagonist SEQ ID NO:15.

FIG. 2 shows a comparison of the change in lung permeability resultingfrom IC formation in response to the treatment described above in thepresence and absence of tight junction antagonist as well as acomparison of the effects of IV versus IT administration of tightjunction antagonist.

FIG. 3 shows the results of varying the dose of SEQ ID NO:15administered with the anti-BSA antibodies delivered IT.

All publications, patents and patent applications mentioned in thisspecification are indicative of the level of skill of those skilled inthe art to which this invention pertains, and are herein incorporated byreference to the same extent as if each individual publication, patentor patent application was specifically and individually indicated to beincorporated by reference.

What is claimed is:
 1. A method for treating acute respiratory distresssyndrome or acute lung injury comprising: administering to a humansubject in need thereof, a composition comprising a peptide tightjunction antagonist, wherein the tight junction antagonist is a peptidehaving the amino acid sequence GGVLVQPG (SEQ ID NO: 15).
 2. The methodaccording to claim 1, wherein the peptide consists of GGVLVQPG (SEQ IDNO: 15).
 3. The method according to claim 1, wherein the composition isadministered in conjunction with an additional therapeutic agent.
 4. Themethod according to claim 3, wherein the composition and the additionaltherapeutic agent are administered simultaneously.
 5. The methodaccording to claim 3, wherein the composition and the additionaltherapeutic agent are not administered simultaneously.
 6. The methodaccording to claim 3, wherein the composition further comprises theadditional therapeutic agent.
 7. The method according to claim 3,wherein the additional therapeutic agent is selected from the groupconsisting of aminosalicylates, corticosteroids, immunomodulators,antibiotics, cytokines, chemokines, and biologic therapeutics.
 8. Themethod according to claim 1, wherein the composition is formulated forpulmonary delivery.
 9. The method of claim 1, wherein the subject hasacute respiratory distress syndrome.
 10. The method of claim 1, whereinthe subject has acute lung injury.
 11. The method of claim 1, whereinthe composition is a pharmaceutical aerosol.
 12. The method of claim 1,wherein the composition is a solution aerosol.
 13. The method of claim1, wherein the composition is a powder aerosol.
 14. The method of claim6, wherein the additional therapeutic agent is an aminosalicylate orcorticosteroid.
 15. The method of claim 1, wherein the composition isadministered intravenously (IV) or intratracheally (IT).